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. 2020 Oct 8;10:16853. doi: 10.1038/s41598-020-73817-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Schematic diagram summarizing the effects of ROCK inhibition and Ca²⁺ on cell sheet cultures. (A) In KCM containing 1.6 mM Ca²⁺ and 10 μM Y-27632, inhibition of ROCK promotes cell proliferation but dephosphorylates MLC to inhibit myosin–actin interactions, resulting in a loose cell sheet that contains defects after harvesting. (B) In KCM containing 3.6 mM Ca²⁺ and 1 μM Y-27632, the elevation of Ca²⁺ recovers MLC phosphorylation in the presence of the ROCK inhibitor, resulting in a cell sheet that shrinks on harvesting without developing defects.